Novel antiseptic derivatives with broad spectrum antimicrobial activity for the impregnation of surfaces

ABSTRACT

The present invention provides novel broad spectrum antiseptic compounds that further have properties that allow them to be coated/impregnated into polymer surfaces. Methods for coating these antiseptic compounds onto medical devices especially in-dwelling medical devices to prevent the growth of pathogens in such devices and hence, to prevent infection to patients via such devices are provided. The invention also provides antiseptics that are useful as general surface disinfectants and sterilizers, fluid disinfectants and biocide preservatives.

[0001] The present invention claims priority to U.S. ProvisionalApplication Serial No. 60/261,447 filed on Jan. 12, 2001 and U.S.Provisional Application Serial No. 60/316,165 filed on Aug. 30, 2001,which are incorporated by reference in their entirety.

BACKGROUND OF THE INVENTION

[0002] 1. Field of the Invention

[0003] The present invention relates generally to the fields ofpreventing infections. More particularly it provides novel broadspectrum antiseptic compositions that further have properties that allowthem to be coated/impregnated into polymer surfaces or used asantiseptics in different applications. The invention provides methodsfor coating these antiseptic compositions onto medical devices such ascatheters, tubes, stents and sutures, to prevent the growth of pathogensin such devices and hence, to prevent infection to patients via suchdevices. In addition, the invention provides novel antiseptics thatcould be used in disinfecting and sterilizing organic and inorganicsurfaces, water and other fluids.

[0004] 2. Description of Related Art

[0005] Most nosocomial infections are caused by the contamination ofmedical devices resulting in serious hospital-acquired infections.Nosocomial pneumonias are the second most common nosocomial infections,and are associated with the highest attributable mortality andmorbidity. Recent data have shown that at least 300,000 episodes ofnosocomial pneumonia occur annually in the United States (OfficialStatement, American Thoracic Society). The attributable mortality ofthis infection is 33%-50%, hence, around 100,000 patients die annuallybecause of nosocomial pneumonia (CDC, 1993; Leu et al., 1989). The riskof nosocomial pneumonia increases 6- to 20-fold from the use ofmechanical ventilation (Official Statement, American Thoracic Society).

[0006] The endotracheal tube is considered a common vehicle forcolonization/contamination leading to nosocomial pneumonia. Theendotracheal tube connects the oropharyngeal environment with thesterile bronchoalveolar space, significantly increasing the risk ofnosocomial pneumonia. Endotracheal tubes are typically constructed ofpolyvinylchloride, which is known to be very difficult to impregnatewith antiseptic or antimicrobial agents. Thus, there are no endotrachealtubes that are impregnated with antibiotics or antiseptics currently inuse.

[0007] Another leading cause of serious nosocomial infections isbloodstream infections. The primary contributors to nosocomialbloodstream infections are vascular catheters. It is estimated thataround 400,000 vascular catheter-related bloodstream infections (CRBSI)occur annually in the United States (Raad, 1998). The attributablemortality of these infections in the intensive care unit (ICU) wasestimated in JAMA in 1994 to be 25% (Reiselman et al., 1994). Hence,these infections are a major cause of morbidity and mortality inhospitalized patients. These catheters are mostly polyurethaneshort-term catheters used in the ICU and long-term silicone cathetersused in cancer/AIDS patients.

[0008] The most frequent causes of nosocomial infections are urinarytract infections (TI), contributing to 34% of all nosocomial infections(Klempner et al., 1998). Nosocomial UTI are usually associated withcontamination of urinary catheters. In addition, nosocomial surgicalwound infections are common complications of surgical procedures,particularly in cancer and immunocompromised patients with devitalizedtissue and decreased immunity. Surgical wound infections contribute to17% of all nosocomial infections (Platt and Bucknall, 1988). Manysurgical wound infections are associated with the contamination ofsutures.

[0009] Antibiotics are strictly antibacterial agents that are usuallyused in treatment of systemic or bloodstream infections and are giventhrough oral, intravenous, subcutaneous, or intramuscular routes toachieve systemic bloodstream levels. Examples include penicillin,cephalosporins, vancomycin, minocycline, and rifampin.

[0010] Antiseptics on the other hand, are antimicrobial agents oftenwith broad spectrum antimicrobial activity against bacteria, fungi orviurses. These agents are used on the skin and external mucosal surfacesusually because of limitations related to absorption, penetration orsystemic toxicity. These agents are not used in the treatment ofbloodstream infections. Examples include chlorhexidine and povidoneiodine.

[0011] Antibiotics and antiseptics have been used to impregnate vascularcatheters. The concern with the use of antibiotics has been thatresistance might develop to antibiotics, preventing their usetherapeutically and systemically in hospitalized patients. Furthermore,the durability of the existing antiseptics has been limited. Forexample, the use of chlorhexidine/silver sulfadiazine on polyurethanesurfaces has had limited effectiveness. Moreover, chlorhexidine/silversulfadiazine impregnating the surface of vascular catheters resulted inlimited activity against gram-negative bacilli, such as Pseudomonas.

[0012] What is needed is an effective antiseptic having broad spectrumactivity against resistant staphylococci, vancomycin-resistantenterococci, resistant Pseudomonas aeruginosa and Candida species, to beused in conjunction with indwelling devices that will inhibit or preventthe nosocomial infections typically associated with the use of theseindwelling devices. It would be further desirable to develop devicesimpregnated with the antiseptic to enhance the resistance to infection.For example, the creation of antiseptic-impregnated catheters wouldprevent organisms from adhering or migrating on catheter surfaces.

SUMMARY OF THE INVENTION

[0013] The present invention overcomes these and other drawbacksinherent in the art by providing novel antiseptic derivatives withbroad-spectrum activity against various microbes including resistantbacteria and fungi. Methods of preparing these antiseptic compounds andmethods for utilizing them are provided.

[0014] Therefore, the invention provides an antiseptic compositioncomprising a basic reagent and a dye. The basic reagent may be bonded tothe dye. In one aspect, the basic reagent and the dye are bondedionically to form the antiseptic compound. In another aspect, the basicreagent and the dye are bonded covalently to form the antisepticcompound. The basic reagent and the dye can be combined in any amount toobtain the antiseptic composition of the invention, however, in aparticular embodiment, an equimolar amount of the basic reagent is addedto the dye solution. The inventors also contemplate that the antisepticcomposition of the invention can be made by combining other amounts ofthe dye and basic reagent for example, one may combine, in molar ratios,1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:25,1:30, 1:35, 1:40, 1:45, 1:50, 1:55, 1:60, 1:65,1:70,1:75, 1:80, 1.85,1:90, 1:95, to 1:99 of either dye:basic reagent or basic reagent:dye.This includes all the intermediate ranges as well, for example itincludes molar ratios such as, 1.1:1, 1.2:1, 1.3:1, 1.4:1, 1.5:1, 1.6:1,1.7:1, 1.8:1, 1.9:1 and the like for other values listed. It alsoincludes the ranges in between these values such as 1.11:1, 1.12:1 andso on. The skilled artisan will therefore recognize that the dye andbasic reagent can be combined in different molar ratio amounts to obtainthe antiseptic composition disclosed and that the invention is thereforenot limited to any particular molar ratio of dye : basic reagent orbasic reagent : dye.

[0015] In certain embodiments, the dye can be a triarylmethane dye, amonoazo dye, a diazo dye, an indigoid dye, a xanthene or a fluoresceindye, an anthraquinone dye, or a quinoline dye. In other specificembodiments, the dye is gentian violet, or crystal violet, ethyl violet,brilliant green, an FD&C dye, or a D&C dye. In one example, the FD&C dyeis Blue No. 1 or Green No. 3. In another example, the triarylmethane dyeis gentian violet. In yet another example, the monoazo dye is FD&CYellow No. 5 or FD&C Yellow No. 6. In still another example, the diazodye is D&C Red No. 17. The indigoid dye, may preferably be FD&C Blue No.2. An example of a xanthene dye is FD&C Red No. 3, of an anthraquinonedye is D&C Green No. 6 And an example of a quinoline dye is D&C YellowNo. 1. In addition, Table 1 provides a list of different dyes that maybe used in this invention. One of skill in the art will recognize thatthese examples are non-limiting and that the antiseptic compounds andcompositions of the present invention can be made using almost any dye.

[0016] A wide variety of basic reagents can be used to form theantiseptic composition. The basic reagents include any nucleophilicspecies which includes all electron donor species. Some of the basicreagents that can be used include a guanidium compound, a biguanide, abipyridine, a phenoxide antiseptic, an alkyl oxide, an aryl oxide, athiol, an aliphatic amine, or an aromatic amine and halides such as F⁻,Br⁻ and I⁻. Some examples of guanidium compounds that may be usedinclude chlorhexidine, alexidine, and hexamidine. One example of abipyridine compound that can be used to synthesize the antiseptics ofthe invention is octenidine. Examples of phenoxide antiseptics usedinclude colofoctol, chloroxylenol, and triclosan.

[0017] The invention also provides an antiseptic compound comprising abasic reagent bound to a dye. In some particular embodiments, the basicreagent and the dye are bound ionically. In other particularembodiments, the basic reagent and the dye are bound covalently. Someexamples of the invention, the antiseptic compound comprisescompositions such as gendine, genlenol, genlosan, or genfoctol.

[0018] In some embodiments, the antiseptic compound provided herein isfurther capable of impregnating and/or coating a surface.

[0019] In some aspects, the surface is composed of a polymer. Examplesof such polymeric surfaces include polyvinyl chloride, polyurethane,polyethylene, silastic elastomers, polytetrafluoroethylene, dacron,collodion, carboethane or nylon. Alternatively, the surface may becomposed of silicone or may be a silk suture. For example, these novelantiseptic derivatives have the potential for serving as impregnators ofmedical device surfaces, such as endotracheal tubes made of polyvinylchloride, vascular catheters made of either polyurethane or silicone,and silk sutures used for suturing surgical wounds.

[0020] In other embodiments, the antiseptic compound can coat and/orimpregnate an organic surface. Examples of organic surfaces include theskin, a mucosal surface, a wound. Examples of wounds are surgicalwounds, trauma wounds, bum wounds and the like.

[0021] In yet other embodiments, the antiseptic compound can coat and/orimpregnate an inorganic surface. Examples of such inorganic surfacesinclude floors, table-tops, counter-tops, surfaces of a hospitalequipment, wheelchair surfaces, etc. Virtually any surface comprising amaterial that is capable of being coated by, impregnated with, absorbingor otherwise retaining the antiseptic compounds of the invention may bedisinfected and/or sterilized using the present antiseptic compounds andtheir compositions. Thus, the antiseptic compound of the invention canbe used to disinfect, sanitize and sterilize a wide variety of surfaces.

[0022] The invention also provides medical devices coated with a basicreagent and a dye. In one aspect the medical devices are coated with abasic reagent and a dye that are ionically bound. In another aspect themedical devices are coated with a basic reagent and a dye that arecovalently bound. Examples of medical devices include endotrachealtubes, a vascular catheter, an urinary catheter, a nephrostomy tube, abiliary stent, a peritoneal catheter, an epidural catheter, a centralnervous system catheter, an orthopedic device, a prosthetic valve, and amedical implant. The vascular catheter may be a central venous catheter,an arterial line, an pulmonary artery catheter, and a peripheral venouscatheter. The central nervous system catheter may be an intraventricularshunt. Other medical devices that can benefit from the present inventioninclude blood exchanging devices, vascular access ports, cardiovascularcatheters, extracorpeal circuits, stents, implantable prostheses,vascular grafts, pumps, heart valves, and cardiovascular sutures, toname a few. Regardless of detailed embodiments, applicability of theinvention should not be considered limited with respect to the type ofmedical device, implant location or materials of construction of thedevice.

[0023] The invention also provides methods for coating a medical devicewith an antiseptic compositions comprising: a) immersing a medicaldevice in a solvent comprising a basic reagent and a dye; b) drying thedevice; and c) washing off excessive composition. In some embodiments,the solvent used to immerse the device can be methylene chloride,methanol, or a combination thereof.

[0024] The invention also provides methods for preventing nosocomialinfections in a subject comprising coating a medical device that thesubject is required to use with a composition comprising an antisepticcompound comprising a basic reagent bound to a dye. The subject can behuman or an animal model.

[0025] The type of nosocomial infection that can be prevented by themethods of this invention include, but are not limited to, pneumonia,bacteremia, fungimia, candidemia, a urinary tract infection, acatheter-exit site infection, and a surgical wound infection.

[0026] The nosocomial infections that can be prevented may be caused bybacteria. In some embodiments the bacteria are drug resistant bacteria.Some non-limiting example of drug resistant bacteria includemethicillin-resistant staphylococci, vancomycin-resistant enterococci,and resistant Pseudomonas aeruginosa.

[0027] The nosocomial infection may be caused by a fungus. In some casesthe fungal agent is a drug resistant fungi. Examples of a drug resistantfungi include members of the Candida species. Other pathogenic organismsthat can cause the nosocomial infections are cited elsewhere in thisspecification and coating devices and surfaces with the antiseptics ofthe present invention can prevent infections by these organisms as well.

[0028] The invention also provides methods for disinfecting and/orsterilizing a surface comprising applying the antiseptic composition ofthe invention to the surface. Examples of surfaces that may bedisinfected and/or sterilized include organic surfaces such as skin,mucosal surfaces, wound surfaces and the like. Other examples includeinorganic surfaces such as floors, table-tops, counter-tops, hospitalequipment, wheel chairs, gauze, cotton. The skilled artisan will realizethat most any surfaces can be disinfected or sterilized by theantiseptic compositions provided herein.

[0029] The invention also provides methods for disinfecting and/orsterilizing a fluid comprising adding a composition comprising a basicreagent and a dye into the fluid. The inventors contemplate thatdifferent types of fluids may be disinfected and some non-limitingexamples include water, such as water in coolers and swimming pools,metal working fluids, and petroleum.

[0030] The invention also provides several novel biocide preservativecompounds comprising the antiseptic compositions described above. Alsoprovided are methods for preserving substances by applying thecompositions of the invention on the substance. A variety of substancesmay be preserved by the biocide preservatives of the invention and theyinclude wood, paint, plastic and paper.

[0031] Thus, the antiseptic compositions of the present invention havebroad uses including use in healthcare by providing sterile medicaldevices and surface sterilization and decontamination, as environmentaldecontaminents, fluid decontaminents and in the industrial world asbiocide preservatives.

[0032] As used herein the specification and claim(s), the words “a” or“an” when used in conjunction with the word “comprising” may mean one ormore.

[0033] As used herein the specification and claim(s), the words “ionicbonding” or “ionically bound” refers to the electrostatic interactionsamong ions which can be formed by the transfer of one or more electronsfrom one atom or group of atoms to another, to create an ionic bondbetween the basic reagent and the dye comprising an antiseptic compound.

[0034] As used herein the specification and claim(s), the words“covalent bonding” or “covalently bound” refers to the chemical bondformed by the sharing of one or more pairs of electrons between thebasic reagent and the dye comprising an antiseptic compound.

[0035] Other objects, features and advantages of the present inventionwill become apparent from the following detailed description. It shouldbe understood, however, that the detailed description and the specificexamples, while indicating preferred embodiments of the invention, aregiven by way of illustration only, since various changes andmodifications within the spirit and scope of the invention will becomeapparent to those skilled in the art from this detailed description.

DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

[0036] A. The Present Invention

[0037] Indwelling catheters and other similar implanted medical devicesare used routinely in hospitals on a diverse group of patients. A commoncause of failure of these medical devices is infection. Pathogens oftenattach to and proliferate in such devices and eventually invade thepatient leading to nosocomial infections. Microorganisms usually migratealong the surfaces of devices to invade sterile environments, such asthe bronchoalveolar space leading to pneumonia, the bloodstream leadingto bacteremia, or the urinary bladder leading to urinary tractinfections.

[0038] The present invention provides a series of novel antisepticcompositions with broad-spectrum activity against various nosocomialmicroorganisms, including resistant bacteria and fungi. For example, theantiseptic compositions are effective against resistant staphylococci,vancomycin-resistant enterococci, resistant Pseudomonas aeruginosa andCandida species. These novel antiseptics also have unique propertiesthat enable penetration/impregnation of various polymers, such aspolyvinyl chloride, polyethylene, silastic elastomers,polytetrafluoroethylene, dacron, collodion, carboethane, nylon, polymersused in the formation of endotracheal tubes, silicone and polyurethanepolymers used in the formation of vascular catheters and surgical silksutures. Thus, they are suitable for coating a wide range of devicesurfaces.

[0039] The inventors demonstrate herein that these novel antisepticsmaintain prolonged antimicrobial activity on device surfaces, and thusmay be used for the entire lifespan of these indwelling devices. This isan improvement over existing coated or impregnated devices where theantimicrobial activity of the device diminishes over time and eventuallydisappears altogether. For example, several prior art patents andpublications describe methods of coating which generated medical deviceswherein the effectiveness of the coating diminishes over time. Afterinsertion of the medical device, the antibiotics and/or antisepticsquickly leach from the surface of the device into the surroundingenvironment. Over a relatively short period of time, the amount ofantibiotics and/or antiseptics present on the surface decreases to apoint where the protection against bacterial and fungal organisms is nolonger effective. Thus, the present invention provides safe antiseptictreated devices wherein the antiseptic coating has a durability that maylast through the life-span of the device. This significantly decreasespatient mortality and morbidity associated with the various nosocomialinfections such as nosocomial pneumonias, nosocomial bacteremias,nosocomial urinary tract infections and nosocomial surgical woundinfections. For example, creation of antiseptic-impregnated/coatedcatheters prevents organisms from adhering to or migrating on cathetersurfaces. Thus, when a pathogenic organism approaches the cathetersurface, it is killed by the antiseptics.

[0040] The invention also provides methods for the synthesis of novelbroad-spectrum antiseptic derivatives. The general method for synthesisof the novel antiseptic derivatives involves the binding of a dye withone or more basic reagents. Different types of dyes and basic reagentscan be used to prepare the antiseptic compounds of this invention.

[0041] The dyes that may be used to synthesize the antiseptic compoundsof the invention include but are not limited to, gentian, or crystalviolet, ethyl violet, brilliant green, etc., and the FD&C dyes such asBlue No. 1 and Green No. 3. In addition, other dyes include thefollowing FD&C and D&C colors: (1) Monoazo dyes such as, but not limitedto, FD&C Yellow No. 5, FD&C Yellow No. 6, (2) Diazo dyes such as, butnot limited to, D&C Red No. 17, (3) Indigoid dyes such as, but notlimited to, FD&C Blue No. 2, (4) Xanthene (Fluorescein) dyes such as,but not limited to, FD&C Red No. 3, (5) Anthraquinone dyes such as, butnot limited to, D&C Green No. 6, (6) Quinoline dyes such as, but notlimited to, D&C Yellow No. 1. An extensive list of dyes and stains thatmay be employed is also provided in Table 1.

[0042] The basic reagents can be alkyl and aryl oxides, thiols,sulfides, phosphorous, aliphatic and aromatic amines, guanidines andhalides such as F⁻, Br⁻ and I⁻. Some examples of the basic reagents thatcan be used include phenoxide antiseptics (such as clofoctol,chloroxylenol, triclosan) or guanidium compounds (such as chlorhexidine,alexidine, hexamidine) or bipyridines (such as octenidines). TABLE 1 TheColor Index (C.I.) Number and/or Chemical Abstracts Service RegistryCAS) Number for Dyes and Stains that may be Employed to Stain MedicalDevices: No. C.I. # CAS # 1 15670 2092-55-9 2 26370 3071-73-6 3 204605850-35-1 4 62130 2666-17-3 5 61585 4474-24-2 6 26360 3351-05-1 7 620586397-02-0 8 42685 3244-88-0 9 61580 6408-57-7 10 15575 5850-86-2 1122870 15792-50-4 12 18050 3734-67-6 13 14900 4787-93-3 14 1807012167-45-2 15 22890 10169-02-5 16 23635 6459-94-5 17 18800 6408-31-7 1818055 4321-69-1 19 18965 6359-98-4 20 18900 6359-91-7 21 2513513390-47-1 22 22910 6375-5-9 23 18850 6359-88-2 24 46005:1 494-38-2 258048-52-0 26 58000 72-48-0 27 3952-78-1 28 61710 6408-63-5 29 4275030586-13-1 30 569-58-4 31 52417-22-8 32 520-10-5 33 48035 3056-93-7 344431-00-9 35 50090 25360-72-9 36 52010 531-55-5 37 61111 12217-43-5 3842500 569-61-9 39 11460 42373-04-6 40 23500 992-59-6 41 298-95-3 4221010 5421-66-9 43 1871-22-3 44 28440 2519-30-4 45 42660 6104-59-2 4627290 5413-75-2 47 24890 3051-11-4 48 76-60-8 49 115-40-2 50 115-39-9 5165005 1328-24-1 52 62055 6408-78-2 53 62125 6424-85-7 54 63010 2861-02-155 13390 3861-73-2 56 26400 3529-01-9 57 15706 12392-64-2 58 615704403-90-1 59 62560 4430-16-4 60 26550 8003-88-1 61 18745 10127-27-2 6214710 5858-39-9 63 17045 6360-07-2 64 15620 1658-56-6 65 18110 6844-74-266 26900 6406-56-0 67 18125 10130-48-0 68 42650 4129-84-4 69 188356359-85-9 70 18890 6359-90-6 71 18950 6372-96-9 72 14170 6408-90-8 7313900 10343-58-5 74 46025 135-49-9 75 12840 61968-76-1 76 636151324-21-6 77 58005 130-22-3 78 14025 584-42-9 79 42080 3486-30-4 8016185 915-67-3 81 42780 82 1668-00-4 83 41000 2465-27-2 84 4381013186-45-3 85 52005 531-53-3 86 51004 33203-82-6 87 11075 94233-04-2 8842510 632-99-5 89 48055 4208-80-4 90 26905 4196-99-0 91 2315-97-1 9221000 10114-58-6 93 16180 5858-33-3 94 42655 6104-58-1 95 81029-05-2 9642040 633-03-4 97 102185-52-4 98 62625-32-5 99 62625-30-3 100 62625-28-9101 14337-53-2 102 76-59-5 103 40070-59-5 104 3147-14-6 105 244102610-05-1 106 43825 1667-99-8 107 16575 548-80-1 108 43820 3564-18-9 10924895 2870-32-8 110 18972 50662-99-2 111 596-27-0 112 2303-01-7 1131733-12-6 114 10510-54-0 115 15970 1934-20-9 116 15391-59-0 117 76-54-0118 4727-50-8 119 54-88-6 120 6473-13-8 121 23655 6420-03-7 122 253802829-43-8 123 27905 5489-77-0 124 13950 10190-68-8 125 29025 3214-47-9126 64500 2475-45-8 127 61500 2475-44-7 128  1005 730-40-5 12931482-56-1 130 11115 3180-81-2 131 11855 2832-40-8 132 26090 6300-37-4133 45400 548-24-3 134 45380 548-26-5 135 15086-94-9 136 14640 3564-14-5137 42090 3844-45-9 138 45430:2 15905-32-5 139 45386 6359-05-3 14076058-33-8 141 23860 314-13-6 142 11160 97-56-3 143 13015 2706-28-7 14411285 6416-57-5 145 45350:1 2321-07-5 146 596-09-8 147 3326-34-9 14851030 1562-85-2 149 1634-82-8 150 3737-95-9 151 165660-27-5 15216574-43-9 153 34722-90-2 154 617-19-6 155 51050 1562-90-9 156 4430-20-0157 14720 3567-69-9 158 16570 4197-07-3 159 11270 532-82-1 160 1810517681-50-4 161 22120 573-58-0 162 2411-89-4 163 62625-31-4 16462625-29-0 165 41830-80-2 166 42555 548-62-9 167 45370:1 596-03-2 168620-45-1 169 45425:1 31395-16-1 170 73688-85-4 171 34140 4399-55-7 17229160 3441-14-3 173 28160 2610-11-9 174 13920 10130-29-7 175 195566537-66-2 176 36900 6409-90-1 177 61505 2475-46-9 178 11080 2581-69-3179 26080 6253-10-7 180 11110 2872-52-8 181 11130 2734-52-3 182 127906439-53-8 183 518-82-1 184 56360-46-4 185 45380:2 15086-94-9 186 146451787-61-7 187 18760 3618-63-1 188 45430 568-63-8 189 1239-45-8 19062758-12-7 191 42600 2390-59-2 192 37190 64071-86-9 193 42053 2353-45-9194 12010 6535-42-8 195 18820 6359-82-6 196 45350 518-47-8 197 3326-32-7198 51649-83-3 199 42085 4680-78-8 200 75290 517-28-2 201 90-33-5 20273000 482-89-3 203 73015 860-22-0 204 12210 4569-88-4 205 110502869-83-2 206 44090 3087-16-9 207 42000 2437-29-8 208 13065 587-98-4 20952041 2516-05-4 210 45385 23391-49-3 211 13025 547-58-0 212 32469-43-5213 14855 3624-68-8 214 11335 6247-27-4 215 11880 6370-46-3 216 113006232-53-7 217 26520 3564-27-0 218 18735 1934-24-3 219 14010 6054-99-5220 44530 5715-76-4 221 11350 131-22-6 222 16255 2611-82-7 223 520306586-05-6 224 7385-67-3 225 74-39-5 226 60760 6409-77-4 227 261204477-79-6 228 16230 1936-15-8 229 15705 2538-85-4 230 19010 10127-05-6231 42045 129-17-9 232 34487-61-1 233 101-75-7 234 11800 1689-82-3 23545410 18472-87-2 236 16680 1058-92-0 237 27190 6226-78-4 238 4900030113-37-2 239 16593-81-0 240 85531-30-2 241 45005 92-32-0 242 5850081-61-8 243 47000 8003-22-3 244 20505 17095-24-8 245 61205 13324-20-4246 17908 25489-36-5 247 635-78-9 248 45170 81-88-9 249 45160 989-38-8250 45440 632-69-9 251 50240 477-73-6 252 61552 6994-46-3 253 7423-31-6254 3599-32-4 255 146-68-9 256 42095 5141-20-8 257 42000:1 510-13-4 258129-16-8 259 52015 61-73-4 260 50206 4569-86-2 261 42590 7114-03-6 26213020 493-52-7 263 11020 60-11-7 264 20110 3564-15-6 265 11875 6247-28-5266 13250 3618-62-0 267 14030 2243-76-7 268 26560 6406-37-7 2696408-91-9 270 14045 6470-98-0 271 20470 1064-48-8 272 50040 553-24-2 27342520 3248-91-7 274 51180 3625-57-8 275 14890 5423-07-4 276 56431-61-9277 61555 2646-15-3 278 26125 1320-06-5 279 15510 633-96-5 280 157115610-64-0 281 12070 6410-10-2 282 143-74-8 283 11000 60-09-3 28416201-96-0 285 975-17-7 286 2768-90-3 287 27195 6226-79-5 288 67627-18-3289 58205 (75410) 81-54-9 290 115-41-3 291 45010 2150-48-3 292 117-92-0293 58050 81-64-1 294 47005 8004-92-0 295 61211 12236-82-7 296 1775712225-82-1 297 61200 2580-78-1 298 123333-76-6 299 45170:1 509-34-2 30013161-28-9 301 43800 603-45-2 302 61554 17354-14-2 303 61565 128-80-3304 12055 842-07-9 305 12140 3118-97-6 306 26105 85-83-6 307 119202051-85-6 308 123359-42-2 309 23647-14-5 310 45100 3520-42-1 311 191401934-21-0 312 108321-10-4 313 62637-91-6 314 6262-21-1 315 632-73-5 31642798-98-1 317 19540 1829-00-1 318 52000 78338-22-4 319 81012-93-3 320123359-43-3 321 12120 2425-85-6 322 23850 72-57-1 323 44045 2580-56-5324 42595 2390-60-5 325 125-31-5 326 16150 3761-53-3 327 135-52-4 32826100 85-86-9 329 26150 4197-25-5 330 26050 6368-72-5 331 68504-35-8 332123333-78-8 333 45220 5873-16-5 334 4430-25-5 335 1301-20-8 336123333-63-1 337 386-17-4 338 4430-24-4 339 1719-71-7 340 49005 2390-54-7341 76-61-9 342 125-20-2 343 52040 92-31-9 344 14270 547-57-9 34514541-90-3 346 44040 2185-86-6 347 45190 6252-76-2 348 63721-83-5 34914936-97-1

[0043] One unique feature of these novel antiseptics is that they do notrequire another vehicle to attach to a surface. The adhesive potentialof the dye makes them self-adhesive to surfaces of devices.

[0044] The antiseptic compound is therefore applied on the surface of adevice by simply immersing the device in the antiseptic solution, airdrying and washing out excessive antiseptic. The self-impregnatingproperty of the dyes such as for example, the triarylmethane dyes,removes the need for another binding agent. This is another feature ofthe composition provided by this invention which is a considerableimprovement over other known compositions. Previously known compositionsrequire other impregnating/coating agents and/or must typically beextruded into the device as it is made. Both these methods are timeconsuming and involve additional steps and techniques. For example, onemethod of coating devices first requires application or absorption of alayer of surfactant, such as tridodecylmethyl ammonium chloride (TDMAC)followed by the antibiotic coating layer, to the surface of the medicaldevice. Another method used to coat surfaces of medical devices withantibiotics involves first coating the selected surfaces withbenzalkonium chloride followed by ionic bonding of the antibioticcomposition (Solomon and Sherertz, 1987; U.S. Pat. No. 4,442,133). Othermethods of coating surfaces of medical devices with antibiotics aretaught in U.S. Pat. No. 4,895,566 (a medical device substrate carrying anegatively charged group having a pH of less than 6 and a cationicantibiotic bound to the negatively charged group); U.S. Pat. No.4,917,686 (antibiotics are dissolved in a swelling agent which isabsorbed into the matrix of the surface material of the medical device);U.S. Pat. No. 4,107,121 (constructing the medical device with ionogenichydrogels, which thereafter absorb or ionically bind antibiotics); U.S.Pat. No. 5,013,306 (laminating an antibiotic to a polymeric surfacelayer of a medical device); and U.S. Pat. No. 4,952,419 (applying a filmof silicone oil to the surface of an implant and then contacting thesilicone film bearing surface with antibiotic powders). Furthermore,most of the methods previously employed to coat the surfaces of medicaldevices use antibiotics such as tetracyclines, penicillins,cephalosporins and the beta-lactam antibiotics. The main drawback withantibiotics is the emergence of resistant strains.

[0045] Thus, the invention provides novel antiseptic derivativecompounds with broad-spectrum antiseptic activity against bacteria andfungi including nosocomial and multidrug-resistant varieties with theadditional ability to impregnate, bind, coat, adhere and/or attach tovarious device surfaces without the assistance of impregnating vehiclessuch as tridodecylmethylammonium chloride (TDMAC). Furthermore, thenovel antiseptic compounds of the invention also have an extendedantimicrobial efficacy that can cover the life of the device.

[0046] One example of the novel broad-spectrum antiseptic derivatives ofthis invention is gendine, which consists of the combination of gentianviolet and chlorhexidine. Gentian violet, on its own, is a goodimpregnating triarylmethane dye. Bhatnager et al., 1993 have shown in anin vitro study that gentian violet alone can be used to impregnate thesurface of CSF silicone shunts and prevent the colonization of S.epidermis on these surfaces. However, after impregnating the surfaces ofvarious polymers, including polyvinylchloride, gentian violet on its ownhas no activity against Pseudomonas aeruginosa, which is the second mostcommon cause of nosocomial pneumonia and the third most common cause ofnosocomial urinary tract infections. Antiseptics such as chlorhexidinecannot attach on their own onto the surfaces of polyvinylchloride tubesor silicone catheters and silk sutures. They require an impregnatingvehicle. Furthermore, on their own they are not highly active againstPseudomonas aeruginosa. On the other hand, upon the binding of gentianviolet with chlorhexidine, the new antiseptic agent synthesized, is apotent and effective broad-spectrum antiseptic and has the additionalability to coat/impregnate various device surfaces. Gendine is unique inits ability to impregnate various device polymers, such aspolyvinylchloride used in the formation of endotracheal tubes, siliconeand polyurethane polymers used in the formation of vascular, as well asperitoneal, epidural, urinary and intraventricular catheters. Inaddition, gendine is able to impregnate the silk sutures used insurgical wounds.

[0047] In addition to Gendine, other antiseptics encompassed by thisinvention are Genlenol and Genfoctol.

[0048] The invention also provides methods to generate a wide variety ofantiseptic medical devices. Some examples include antisepticendotracheal tubes, antiseptic vascular catheters, including centralvenous catheters, arterial lines, pulmonary artery catheters, andperipheral venous catheters, antiseptic urinary catheters, antisepticnephrostomy tubes, antiseptic biliary stents, antiseptic peritonealcatheters, antiseptic epidural catheters, antiseptic central nervoussystem catheters, including intraventricular shuts and devices,antiseptic prosthetic valves, orthopedic implants and antisepticsutures.

[0049] B. Pathogens

[0050] The nosocomial bacterial infections result in diseases such asbacteremia, pneumonia, meningitis, osteomyelitis, endocarditis,sinusitis, arthritis, urinary tract infections, tetanus, gangrene,colitis, acute gastroenteritis, bronchitis, and a variety of abscesses,and opportunistic infections. Bacterial pathogens include Gram-positivecocci such as Staphylococcus aureus, coagulase negative staphylocci suchas Staphylococcus epidermis, Streptococcus pyogenes (group A),Streptococcus spp. (viridans group), Streptococcus agalactiae (group B),S. bovis, Streptococcus (anaerobic species), Streptococcus pneumoniae,and Enterococcus spp.; Gram-negative cocci such as Neisseriagonorrhoeae, Neisseria meningitidis, and Branhamella catarrhalis;Gram-positive bacilli such as Bacillus anthracis, Corynebacteriumdiphtheriae and Corynebacterium species which are diptheroids (aerobicand anerobic), Listeria monocytogenes, Clostridium tetani, Clostridiumdifficile, Escherichia coli, Enterobacter species, Proteus mirablis andother spp., Pseudomonas aeruginosa, Klebsiella pneumoniae, Salmonella,Shigella, Serratia, and Campylobacterjejuni. The antibiotic resistantbacteria that can be killed by the antiseptic coated devices of thepresent invention include Staphylococci (methicillin-resistant strains),vancomycin-resistant enterococci (Enterococcus faecium), and resistantPseudomonas aeruginosa.

[0051] Fungal infections that may be prevented include fungal infections(mycoses), which may be cutaneous, subcutaneous, or systemic.Superficial mycoses include tinea capitis, tinea corporis, tinea pedis,onychomycosis, perionychomycosis, pityriasis versicolor, oral thrush,and other candidoses such as vaginal, respiratory tract, biliary,eosophageal, and urinary tract candidoses. Systemic mycoses includesystemic and mucocutaneous candidosis, cryptococcosis, aspergillosis,mucormycosis (phycomycosis), paracoccidioidomycosis, North Americanblastomycosis, histoplasmosis, coccidioidomycosis, and sporotrichosis.Fungal infections include opportunistic fungal infections, particularlyin immunocompromised patients such as those with AIDS. Fungal infectionscontribute to meningitis and pulmonary or respiratory tract diseases.Other pathogenic organisms that may be prevented from causing theinfections include dermatophytes (Microsporum canis and other M spp.;and Trichophyton spp. such as T. rubrum, and T. mentagrophytes), yeasts(e.g., Candida albicans, C. Parapsilosis, C. glabrata, C. Tropicalis, orother Candida species including drug resistant Candida species),Torulopsis glabrata, Epidermophytonfloccosum, Malassezia fuurfur(Pityropsporon orbiculare, or P. ovale), Cryptococcus neoformans,Aspergillus fumigatus, and other Aspergillus spp., Zygomycetes(Rhizopus, Mucor), hyalohyphomycosis (Fusarium Spp.), Paracoccidioidesbrasiliensis, Blastomyces dermatitides, Histoplasma capsulatum,Coccidioides immitis, and Sporothrix schenckii. Fungal infectionsinclude Cladosporium cucumerinum, Epidermophyton floccosum, andMicrospermum ypseum.

C. EXAMPLES

[0052] The following examples are included to demonstrate preferredembodiments of the invention. It should be appreciated by those of skillin the art that the techniques disclosed in the examples which followrepresent techniques discovered by the inventor to function well in thepractice of the invention, and thus can be considered to constitutepreferred modes for its practice. However, those of skill in the artshould, in light of the present disclosure, appreciate that many changescan be made in the specific embodiments which are disclosed and stillobtain a like or similar result without departing from the spirit andscope of the invention.

Example 1 Synthesis of Gendine and Impregnation of Devices

[0053] Impregnation Procedure

[0054] The general procedure involves, and when applicable, priorpreparation of the basic reagent (such as chlorhexidine) in anhydroussolvent, addition of the basic reagent to a solution of a dye (such asGentian violet) in anhydrous solvent (or addition of the dye to thebasic solution), stirring the resulting mixture for 30-90 minutes atambient conditions, evaporating the solvent also under ambientconditions, and finally dissolution of the residue prior toimpregnation. The following procedure illustrates impregnation withGendine, an example for employing a basic guanidium derivative (e.g.,chlorhexidine) and triarylmethane dye (e.g., Gentian violet).

[0055] Potassium tert-butoxide in THF, 7.35 ml of 1M solution, was addedto a solution of CHX diacetate, 1.533 g; 2.45 mmol in 35 ml THF. Theresulting heterogeneous solution was stirred for 20 minutes, then addedto a solution GV, 1.0 g; 2.45 mmol, in 30 ml THF (GV used as an exampleof Triarylmethane Dye). The mixture was stirred at ambient conditionsfor 1 hour, then placed under the hood overnight to evaporate thesolvent. The resulting residue was dissolved in 30 ml DCM (or MeOH).When applicable, the base (such as neutral form of chlorhexidine) isadded to a stirring solution of dye (such as GV) in DCM and theresulting mixture is stirred for at least 1 h. With anionic dyes,dissolution is achieved with the addition of at least one equivalent ofa quaternary amine (such as tetraethylammonium) prior to addition of thebase. One-centimeter device segments were immersed in the DCM solutionfor the appropriate period, generally PVC and PU for 10 minutes;Silicone (Si) and Silk Suture for 2 hours. The devices were removed fromthe solution, and traces of solution were removed from the lumen whenapplicable. The impregnated devices were placed under the hood to dryfor at least 4 hours, preferably over night, then washed with distilledwater until the washings were colorless or very faint, and finallyplaced under an aseptic hood to dry under ambient conditions for atleast 4 hours, preferably overnight.

[0056] In Vitro Antimicrobial Activity

[0057] The antimicrobial activity of impregnated catheters was evaluatedin duplicate by a modified Kirby-Bauer technique. BBL Mueller Hinton IIagar plates (obtained from Fisher Scientific) were inoculated with 0.5McFarland of the appropriate microorganism (hospital isolates from theMD Anderson Cancer Center). Then 10 mm segments of impregnated deviceswere embedded in the inoculated plates and placed in an incubator atabout 37° C. for at least 18 hours. Zones of inhibition were measuredperpendicular to the long axis of the device. Bioactivity against yeastconstituted of two zones; a large outer zone of partial growthinhibition, and an inner smaller zone of complete inhibition. This laterzone is reported herein.

[0058] Results and Discussion

[0059] Tables 2 and 3 illustrate zones of inhibition obtained forGendine-impregnated devices: TABLE 2 Endotracheal PVC Tubes, (7.0 mmI.D.) Zones of Inhibition (in mm) obtained for Reagent in DCM MRSA₂₀₆₆PS₄₂₀₅ C. Parap. ₁₋₁₀₀₋₀₀₂₂ GV^(†) 25:25 0:0 27:27 CHX^(††, §) 0:0 0:00:0 GN, 1^(st) trial^(†) 28:28 21:21 27:28 GN, 2^(nd) trial^(‡) 28:2922:23 27:27

[0060] TABLE 3 Endotracheal PVC Tubes, (7.0 mm I.D.) Zones of Inhibition(in mm) obtained for Reagent in MeOH MRSA₂₀₆₆ PS₄₂₀₅ C. Parap.₁₋₁₀₀₋₀₀₂₂ GV^(†) 20:21 0:0 18:19 CHX^(†) 0:0 0:0 0:0 GN^(†) 24:25 13:1323:23

[0061] As shown in Tables 2 and 3, endotracheal PVC tubes impregnatedwith Gendine (GN) are far more effective against all organisms whencompared with those impregnated with CHX, and more effective than PVCtubes impregnated with GV against Pseudomonas aeruginosa. TABLE 4 Doublelumen 10.0 FR-Cook Silicone Catheter Zones of Inhibition (in mm)obtained for Reagent in DCM MRSA₂₀₆₆ PS₄₂₀₅ C. Parap. ₁₋₁₀₀₋₀₀₂₂ GV^(†)6:7 0:0 0:0 CHX^(†, §) 0:0 0:0 0:0 GN, 1^(st) trial^(†) 18:19 11:1219:19 GN, 2^(nd) trial^(†) 19:19 (19:20)^(‡) 10:11 (12:13)^(‡) 18:18(24:25)^(‡)

[0062] Again data in Table 4 shows how silicone catheters impregnatedwith GN are more effective in inhibiting MRSA, PS and C. parapsilosisthan catheters impregnated with either GV or CHX. TABLE 5 Double lumen10.0 FR-Polyurethane Catheter Zones of Inhibition (in mm) obtained forReagent in DCM MRSA₂₀₆₆ PS₄₂₀₅ C. Parap. ₁₋₁₀₀₋₀₀₂₂ GV^(†) 22:22 0:022:23 CHX^(††, §) 17:17 10:10 15:15 GN, 1^(st) trial^(†) 21:21 13:1426:27 GN, 2^(nd) trial^(‡) 22:22 15:15 22:23

[0063] Similarly, Table 5 indicates that PU catheters impregnated withGN are more effective than PU catheter impregnated with GV in inhibitingPseudomonas aeruginosa, and more effective than PU catheters impregnatedwith CHX against all three organisms, MRSA, PS and C. parapsilosis.TABLE 6 Silk Sutures Zones of Inhibition (in mm) obtained for ReagentMRSA₂₀₆₆ PS₄₂₀₅ C. Parap. ₁₋₁₀₀₋₀₀₂₂ GV^(†) 8:8 0:0 0:0 CHX^(†, §) 0:00:0 0:0 GN, 1^(st) trial^(†) 17:17 3:5 21:21 GN, 2^(nd) trial^(†) 15:154:4 14:14

[0064] Table 6 as well shows that silk sutures coated or impregnatedwith GN are significantly more effective in inhibiting MRSA, PS and C.parapsilosis than sutures coated with either GV or CHX.

[0065] In addition to the simplicity of the impregnation procedure, andthe availability of the requisite reagents, one unique feature ofgendine-impregnated devices is its broad spectrum activity, not onlyagainst the worldwide problematic gram-positive MRSA, which hasincreased in frequency at an alarming rate as a cause of device-relatedinfections, but also against Pseudomonas aeruginosa, an increasinglyprevalent opportunistic human pathogen, and the most commongram-negative bacterium found in nosocomial infections. They areintrinsically more resistant than gram-positive bacteria to manyantiseptics, particularly when present in a biofilm or when associatedwith a device infection (Platt et al., 1988).

[0066] For PVC and silicone (Si) devices, this invention shows noactivity against Pseudomonas aeruginosa, when impregnated with eitherthe immobilizing dye (gentian violet) or CHX alone. All devicesimpregnated with GN exhibit fair to good activity against Pseudomonasaeruginosa. This is important especially since Pseudomonas aeruginosa isresponsible for 16% of nosocomial pneumonia cases (and is considered bythe Centers for Disease Control as the second most common cause ofnosocomial ventilator associated pneumonia), 12% of nosocomial urinarytract infections, 8% of surgical wound infections, and 10% of nosocomialbloodstream infections (Van Delden and Iglewski, 1998).

[0067] Staphylococcal resistance to antiseptics are known worldwide(Russel A. D., 1997). In addition to CHX, low-level resistance to threeantiseptics (acriflavin, benzalkonium chloride, and hexamidinediisethionate is documented (Reverdy et al., 1992; Townsend et al.,1985; Heir et al., 1995). The present study reveals that allGN-impregnated devices, including sutures, exhibit significant biocidalactivity against methicillin-resistant staphylococci. This finding isextremely important in light of the fact that methicillin-resistantstaphylococci (MRSA and MRSE) are the leading causes of device-relatedinfections, including vascular catheter-related bacteremia and surgicalwound infections. In addition S. aureus is one of the leading causes ofnosocomial pneumonia (Klempner et al., 1998).

[0068] The effectiveness of gendine-impregnated devices against Candidais no less noteworthy. As revealed from this study, silicone catheterand suture impregnated with GN exhibit fair to good activity against C.Parapsilosis, which is not the case for either GV or CHX-impregnateddevices. Catheter-related candidemia is now the third leading cause ofvascular catheter-related bloodstream infections (Raad et al., 1992). Inaddition, candidemia in severely immunocompromised patients (i.e., HIV,bone-marrow recipients and leukemia patients) is an important cause formorbidity and mortality and catheters are a major source for thisinfection (Tumbarello et al., 1998; Gonzalez et al., 1996; Lecciones etal., 1992; Wey et al., 1989). The known chlorhexidine-sulfadiazineimpregnated catheters and the minocycline-rifampin impregnated cathetersdo not have significant prophylactic effect against fungi (Tacconelli etal., 1997; Raad et al., 1997).

[0069] Other GV Preparations

[0070] Antiseptics chloroxylenol [p-chloro-m-xylenol;4-chloro-3,5-dimethylxylenol (PCMX)], Clofoctol[α-2,4-dichlorophenly)-4-(1,1,3,3-tetramethylbutyl)-o-cresol (CFTL), andTriclosan [2,4,4′-trichloro-2′hydroxydiphenyl ether] (TLS) are three ofthe phenolic antiseptic reagents included in this study. The firstdisinfectant is the first halophenol employed in many antiseptic anddisinfectant formulations. Neither the neutral form of PCMX nor itssodium salt could produce in vitro zones of inhibition on their ownthrough coating or impregnating the catheter devices and sutures.However, when the salt is reacted with GV (as an example of atriarylmethane dye), the resulting products (new products such asGenlenol, Genlosan and Genfoctol) are immobilized on the devicesproducing large in vitro zones of inhibition against various nosocomialpathogens. Meanwhile, Genlenol (GV⁺.PCMX⁻) not only increased the zoneagainst MRSA considerably for the silicone catheter, but also produced alarge zone against C. parapsilosis. Similar results were observed forsilk sutures. Improvements in the zones of inhibition against C.parapsilosis are also observed for PVC tubes and PVC catheters, and to alarge extent against Alcaligenes faecalis (a gram negative bacillaryorganism) for the PVC tubes. These results are summarized in Tables7-10. Similar trends were observed for Genfoctol (GV⁺.CFTL⁻), but withmuch larger zones for sutures against MRSA and C. parapsilosis. TABLE 7Zones of Inhibition (in mm) imparted by endotracheal^(f) PVC tubes Zonesof Inhibition (in mm) against Alcaligenes Reagent MRSA₂₀₆₆ faecalis ₃₆₈₁C. Parap. ₁₋₁₀₀₋₀₀₂₂ GV^(†) 25:25 18:18 27:27 PCMX 0:0 0:0 0:0GV⁺.PCMX⁻* 27:27 27:28 34:34

[0071] TABLE 8 Zones of Inhibition (in mm) imparted by Siliconecatheters^(f) Zones of Inhibition (in mm) against Alcaligenes ReagentMRSA₂₀₆₆ faecalis ₃₆₈₁ C. Parap. ₁₋₁₀₀₋₀₀₂₂ GV^(†) 6:7 0:0 0:0 PCMX^(‡)0:0 0:0 0:0 GV⁺.PCMX⁻* 16:16 0:0 16:16 CFTL^(§) 0:0 0:0 0:0GV⁺.CFTL^(−II) 20:20 10:10 28:29

[0072] TABLE 9 Zones of Inhibition (in mm) imparted by Polyurethanecatheters^(f) Zones of Inhibition (in mm) against Alcaligenes ReagentMRSA₂₀₆₆ faecalis ₃₆₈₁ C. Parap. ₁₋₁₀₀₋₀₀₂₂ GV^(†) 22:22 18:18 22:23PCMX^(‡) 0:0 0:0 0:0 GV⁺.PCMX⁻* 24:24 18:20 31:31 CFTL^(§) 0:0 0:0 0:0GV⁺.CFTL^(−II) 23:23 15:17 30:32

[0073] TABLE 10 Zones of Inhibition Imparted by Silk Sutures Zones ofInhibition (in mm) against Reagent MRSA₂₀₆₆ C. Parap. ₁₋₁₀₀₋₀₀₂₂ GV^(†)8:8 0:0 PCMX^(‡) 0:0 0:0 GV⁺.PCMX⁻* 11:11 5:5 CFTL^(§) 0:0 0:0GV⁺.CFTL^(−II) 17:17 15:16

[0074] In general, many other gentian violet basic preparationssignificantly affect the efficacy and biocidal activity of coatedsutures and silicone-impregnated catheters against MRSA and CParapsilosis. Some examples are shown below in Table 11. TABLE 11 Zonesof Inhibition Imparted by Silicone Catheters and Silk Sutures Zones ofInhibition (in mm) against MRSA₂₀₆₆ C. Parap. ₁₋₁₀₀₋₀₀₂₂ Si- Silk SilkReagent catheter Suture Si-catheter Suture GV^(†) 6:7 8:8 0:0 0:0 GV⁺OH⁻18:19 17:17 19:19 13:14 GV^(†)OCH₃ ⁻ 18:19 15:15 25:25 10:13GV^(†)Glycerin⁻ 19:20 13:14 18:18 9:9 GV^(†)HEDTA^(−4‡) 13:13 7:7  9:134:5 GV^(†)TCSA^(−§) 14:14 12:13 11:11 10:10 GV^(†)MBT^(−II) 13:13 8:812:12 0:0

[0075] The above data clearly demonstrate significant improvement in thebiocidal activity of impregnated or coated silicone catheters and silksutures by the antiseptic derivatives of the invention.

Example 2 Clinical Trials

[0076] The antiseptic devices of the invention pose no significant risk.Hence, preclinical studies (animal studies) may not be required. Thissection is concerned with the development of human treatment protocolsusing the antiseptic medical devices of the present invention.

[0077] The various elements of conducting a clinical trial, includingpatient treatment and monitoring, will be known to those of skill in theart in light of the present disclosure. The following information isbeing presented as a general guideline for use in establishing the useof the antiseptic medical devices.

[0078] Candidates will be patients who are seriously ill and arerequired to use a medical device such as those described in the sectionsabove. The medical devices in these cases will be treated with Gendine,Genelol, Genfoctol, Genlosan or other antiseptic derivatives that can besynthesized by the methods provided herein, and the patients will bemonitored for the occurrence of nosocomial infections.

[0079] To monitor the development of infections and to evaluate theefficacy of the antiseptic coated/impregnated medical devices inpreventing the spread of infectious agents through the devices it iscontemplated that the patients will be examined for appropriate testsevery month. Tests that will be used to monitor the effectiveness of thetreated medical device include: physical exam, X-ray, blood work andother clinical laboratory methodologies used to detect pathogens in thepatients and also methods to detect presence of pathogens in the medicaldevice. Described below is a study guideline for patients using centralvenous catheters.

[0080] A. Efficacy of Central Venous Catheters Coated with Antisepticsof the Invention

[0081] Patient Eligibility.

[0082] Patients will be recruited from intensive care units, bone marrowtransplant and melanoma services and other hospital divisions wherecatheters are used routinely on inpatients. Patients who require a newinsertion of a central venous catheter (CVC) and have none of theexclusion criteria will be approached to obtain informed consent. Theexclusion criteria are the following:

[0083] 1. Age<18years

[0084] 2. Dermatitis over catheter insertion site

[0085] 3. Pregnancy

[0086] 4. Allergy to chlorhexidine or gentian violet

[0087] 5. Expected duration of catheter placement <3 days

[0088] 6. Inability to obtain informed consent

[0089] The eligible consenting patient will be informed that thecatheter to be inserted has either been coated with an antisepticcompound (for example Gendine) or has not been coated, but the subjectwill not be informed as to whether the specific catheter to be insertedcontains the compound.

[0090] Each female with child bearing potential will have a urine sampleprior to catheter placement to test for pregnancy (if appropriate).

[0091] Catheter Insertion.

[0092] Catheters will be inserted into a subclavian vein or internaljugular vein using gown, mask, sterile gloves and full sterile drapes.Skin will be prepped using povidone iodine allowing 1 minute of exposuretime. After insertion, the catheter will be secured to the skin usingtape and the skin puncture site will be covered with povidone-iodineointment. Then, the insertion site and the surrounding area will becovered with sterile gauze and taped securely.

[0093] Catheter Maintenance.

[0094] Catheters will be inspected every 72 hrs for evidence of siteinfection (erythema around catheter, purulent drainage, swellingtenderness over catheter). Every 72 hrs (or sooner if necessary) thedressing will be removed and the exit site will be re-prepped withpovidone-iodine. All fluids, medications, etc. administered through eachlumen will be documented.

[0095] Catheter Types.

[0096] The inventors contemplate using different types of catheters. Forexample, control catheters consist of triple lumen polyurethanecatheters and single lumen polyurethane catheters will be tested amongseveral others. The test catheters will be identical to the controlcatheters in appearance, but they will be coated with the antiseptics ofthe invention, for example, Gendine.

[0097] Trial Design.

[0098] The trial is a prospective randomized design. The patient, thehealth care worker inserting the catheter, the microbiologist culturingthe catheter, and the evaluator will be blinded as to whether thecatheter is coated or not coated with the antiseptics of the invention.They will, however, be identifiable by an assigned code number. Afterinformed consent has been obtained a catheter will be pulled out of abox containing 6 test and control catheter placement trays. The boxeswill consist of either triple or single lumen catheters and will belabeled as such. The trays will be placed in the boxes such that testand control catheters will alternate from top to bottom. Each box willcontain 3 test and 3 control catheters. The unique identification numberof the catheter will be recorded and will be included with the dataanalysis. Both the investigators and the patients will be blinded to thecatheter identity throughout the study.

[0099] Statistical Considerations.

[0100] Assuming a conservative baseline colonization and/or infectionrate of at least 20% for central venous catheters, randomizing 75patients to each arm would allow one to detect a change incatheter-related infection rates from 20% to 5% one sided significanceand 80% power. If, after entering 150 patients, the infection rate inthe test arm has dropped by 50% (that is from 20% to 10%) then the studywill be expanded to include 400 patients (200 in each arm). Using theselection criteria described above, the inventors estimate that theywill test about 40 patients each month. Aiming for a total of 150evaluable patients the study will be completed in approximately 6months.

[0101] Termination of Study.

[0102] Patients will be kept on study until the catheter is removed. Theindication for catheter removal will be documented for each catheter.These include but are not limited to:

[0103] 1. Catheter no longer needed

[0104] 2. Catheter leaks

[0105] 3. Bleeding around catheter

[0106] 4. Catheter thrombosis

[0107] 5. Catheter insertion site infection or sepsis

[0108] 6. Positive blood cultures that are thought to be clinicallysignificant (i.e. associated fever, increased WBC) and no other site ofinfection is identifiable.

[0109] When the patient becomes febrile, blood will be withdrawnsimultaneously through the lumen of the catheter and peripheral vein forquantitative blood culture. At the time of catheter removal, thecatheter will be removed under aseptic conditions and the tip andintracutaneous segments saved for culturing using the roll plate andsonication quantitative catheter culture technique. At the time ofremoval each lumen will be marked as to its prior use(hyperalimentation).

[0110] Patient Evaluation

[0111]1. Pre-Insertion Evaluation.

[0112] Pertinent history will be taken and physical examination will bedone regarding inclusion and exclusion criteria. Demographic data aswell as details pertaining to underlying malignancy, treatment andantimicrobial treatment (including antimicrobial prophylaxis forinfections in general in patients with hematologic malignancies) will berecorded. Investigational nature of study will be explained and informedconsent will be obtained from patient. Pregnancy tests (serum or urine)will be obtained on all female patients with child bearing potential. Ifthe test is positive, the patient will be excluded.

[0113] Initial catheterization procedure details will be recordedincluding catheter type, site and date of placement; difficulty ofinsertion, and complications if any. The difficulty of insertion will bedetermined by noting the following (a) number of attempts to insert thecatheter (b) time spent during insertion (c) malpositioning andrepositioning of a catheter.

[0114] 2. Post-Insertion Evaluation.

[0115] All patients will be monitored until the catheter is removed.Catheter site evaluation will be undertaken every 72 hrs with the changeof dressing. Special attention will be given to erythema, infiltration,pain, tenderness, swelling, suppuration, palpable cord in vessel, tissuewarmth, lymphangitis or phlebitis. Details pertaining to chemotherapy,antineoplastic and antimicrobials, will be recorded. Catheter usage asfor agents that might cause sclerosis of the vessel involved,hyperalimentation, blood and blood products administration, and drawingof blood will be noted. The catheter insertion site will be recorded onevery patient. In addition, events of repositioning the catheter afterdisplacement will be recorded. Microbiologic evaluation of insertionsite will be undertaken in the form of site cultures if suppuration ispresent. If catheter related septicemia is suspected, blood cultureswill be drawn simultaneously through catheter and by peripheralvenipuncture. Another set of cultures will be drawn 24 hours later. Ifthrombophlebitis is suspected venous flow study of involved vessel willbe done. If line related infection is suspected (including in patientswith fever of unknown origin) or septicemia is documented, catheter willbe changed over guide wire and distal as well as the proximal 5-7 cm ofthe catheter will be evaluated for semiquantitative cultures by theinventors. The purpose of this procedure is diagnostic and nottherapeutic. It will attempt to make a definitive diagnosis of catheterrelated infection by isolating the organism from the catheter usingquantitative techniques.

[0116] 3. End of Evaluation.

[0117] When it is decided to withdraw the line, the catheter will beevaluated by the inventors for quantitative cultures. In addition, aquantitative blood culture will be drawn through the CVC lumen if thelumen is patent and peripherally if the patient is febrile.

[0118] Catheter Assessment

[0119] Definitions:

[0120] 1. Catheter tunnel infection: Either the proximal and/or thedistal catheter segments growing >15 colonies by the roll-plate culturetechnique or >100 colonies by the sonication culture technique.

[0121] 2. Catheter exit site infection: development of lymphangitis,purulence or two of the following: erythema, tenderness, induration orwarmth.

[0122] 3. Catheter related septicemia: Recovery of same organism fromcatheter segment and blood without any other identifiable source for thesepticemia. The catheter should grow at least 15 colonies of theorganism by roll plate or at least 100 colonies by sonication. Thepatient should have clinical manifestation of sepsis (fever, chills orsudden hypotension).

[0123] 4. Catheter-related infection: any of the conditions definedabove would be considered as catheter-related infection.

[0124] Success will be measured if there is no catheter relatedinfection and failure will be indicated by the presence of a catheterrelated infection.

[0125] Adverse Reactions.

[0126] All patients will be monitored for an unexpected adverse reaction(e.g. increased inflammation, phlebitis) associated with the coatedcatheter, using a statistical sequential test method. The study will bestopped if major adverse reaction is identified. Otherwise, the studywill continue until 75 patients in each group have been enrolled.

[0127] All of the compositions and/or methods disclosed and claimedherein can be made and executed without undue experimentation in lightof the present disclosure. While the compositions and methods of thisinvention have been described in terms of preferred embodiments, it willbe apparent to those of skill in the art that variations may be appliedto the compositions and/or methods and in the steps or in the sequenceof steps of the method described herein without departing from theconcept, spirit and scope of the invention. More specifically, it willbe apparent that certain agents which are both chemically andphysiologically related may be substituted for the agents describedherein while the same or similar results would be achieved. All suchsimilar substitutes and modifications apparent to those skilled in theart are deemed to be within the spirit, scope and concept of theinvention as defined by the appended claims.

REFERENCES

[0128] The following references, to the extent that they provideexemplary procedural or other details supplementary to those set forthherein, are specifically incorporated herein by reference.

[0129] American Thoracic Society. Official Statement. Hospital-acquiredpneumonia in adults: diagnosis, assessment of severity, initialantimicrobial therapy and preventative strategies. A consensusstatement. Respir. Crit. Care Med., 153:1711-1725, 1996.

[0130] Bhatnager, Sundaram, Studies on antibacterial properties ofgentian violet impregnated silastic, Indian J. Med. Res., [A]97:206-208,1993.

[0131] Centers for Disease Control and Prevention. Morbidity andMortality Weekly Report CDC Surveillance, 46:891, 1993.

[0132] Delden and Iglewski, Cell-to-cell signaling and Pseudomonasaeruginosa infections, Emerging Infectious Diseases, 4:551-560, 1998.

[0133] Gonzalez, Venzon, Lee, Mueller, Pizzo, Walsh, Risk factors forfungemia in children infected with human immunodeficiency virus: a casecontrol study, Clin. Infect. Dis., 23:515-521, 1996.

[0134] Heir, Sundheim, Holck, Resistance to quaternary ammoniumcompounds in Staphylococcus spp. Isolated from the food industry andnucleotide sequence fo the resistance plasmid pST827, J. Appl.Bacteriol., 79:149-156, 1995.

[0135] Klempner, Lorber, Bartlett, Hospital infections and health-careepidemiology, In: INFECTIOUS DISEASES: MEDICAL KNOWLEDGE SELF-ASSESSMENTPROGRAM, 2^(ND) EDITION, American College of Physicians, Philadelphia,Pa., pp. 210, 1998.

[0136] Lecciones, Lee, Navarro, Vascular catheter-associated fungemia inpatients with cancer: analysis of 155 episodes, Clin. Infect. Dis.14:875-883, 1992.

[0137] Leu, Kaiser, Mori, Hospital-acquired pneumonia: attributablemortality and morbidity, Am. J. Epidemiol., 129:1258-1267, 1989.

[0138] Platt, Bucknall, MIC tests are not suitable for assessingantiseptic handwashes, J. Hosp. Infect., 11:396-397, 1988.

[0139] Raad, Bodey, Infectious complications of indwelling vascularcatheters, Clin. Infect. Dis. 15:197-210, 1992.

[0140] Raad, Darouiche, Dupuis, Central venous catheters coated withminocycline and rifampin for the prevention of catheter-relatedcolonization and bloodstream infections: a randomized, double-blindtrial, Ann. Intern. Med, 127:267-274, 1997.

[0141] Raad, Intravascular-catheter-related infections, Lancet,351:893-898, 1998.

[0142] Reiselman, Tarara, Wenzel, Nosocomial bloodstream infections inthe critically ill, JAMA, 272:1578-1601, 1994.

[0143] Reverdy, Bes, Nervi, Martra, Fleurette, Activity of fourantiseptics (acriflavin, benzalkonium chloride, chlorhexidinedigluconate and hexamidine di-isethionate) and of ethidium bromide on392 strains representing 26 Staphylococcus species, Med. Microbiol.Lett., 1:56-63, 1992.

[0144] Russell, Plasmids and bacterial resistance to biocides, J. App.Microbiol., 82:157-161, 1997.

[0145] Solomon, D. D. and Sherertz, R. J., J. Controlled Release,6:343-352, 1987.

[0146] Tacconelli, Tumbarello, Pittiruti, Central venouscatheter-related sepsis in a cohort of 366 hospitalized patients, Eur.J. Clin. Microbiol. Infect. Dis., 16:203-209,1997.

[0147] Townsend, Ashdown, Momoh, Grubb, Distribution of plasmid-bornresistance to nucleic acid binging compounds in methicillin resistantStaphylococcus aureus, J. Antimicrob. Chemother. 15:417-434, 1985.

[0148] Tumbarello, Tacconelli, Donati, Nosocomial bloodstream infectionsin HIV-infected patients: attributable mortality and extension ofhospital stay, J. Acquir. Immun. Defic. Syndr. Hum. Retrovirol.,19:490-497, 1998.

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What is claimed is:
 1. An antiseptic composition comprising a basicreagent and a dye.
 2. The antiseptic composition of claim 1, wherein abasic reagent and a dye are bonded.
 3. The antiseptic composition ofclaim 2, wherein a basic reagent and a dye are linked by ionic bonding.4. The antiseptic composition of claim 2, wherein a basic reagent and adye are linked by covalent bonding.
 5. The antiseptic composition ofclaim 1, wherein the dye is a triarylmethane dye.
 6. The antisepticcomposition of claim 1, wherein the dye is a monoazo dye.
 7. Theantiseptic composition of claim 1, wherein the dye is a diazo dye. 8.The antiseptic composition of claim 1, wherein the dye is an indigoiddye.
 9. The antiseptic composition of claim 1, wherein the dye is axanthene dye.
 10. The antiseptic composition of claim 1, wherein the dyeis an anthraquinone dye.
 11. The antiseptic composition of claim 1,wherein the dye is a quinoline dye.
 12. The antiseptic composition ofclaim 1, wherein the dye is gentian violet or crystal viol et, ethylviolet, brilliant green, an FD&C dye, or a D&C dye.
 13. The antisepticcomposition of claim 12, wherein the FD&C dye is Blue No. 1 or Green No.3.
 14. The antiseptic composition of claim 5, wherein the triarylmethanedye is gentian violet.
 15. The antiseptic composition of claim 6,wherein the monoazo dye is FD&C Yellow No. 5 or FD&C Yellow No.
 6. 16.The antiseptic composition of claim 7, wherein the diazo dye is D&C RedNo.
 17. 17. The antiseptic composition of claim 8, wherein the indigoiddye is FD&C Blue No.
 2. 18. The antiseptic composition of claim 9,wherein the xanthene dye is FD&C Red No.
 3. 19. The antisepticcomposition of claim 10, wherein the anthraquinone dye is D&C Green No.6.
 20. The antiseptic composition of claim 11, wherein the quinoline dyeis D&C Yellow No.
 1. 21. The antiseptic composition of claim 1, whereinthe basic reagent is a guanidium compound, a biguanide, a bipyridine, aphenoxide antiseptic, an alkyl oxide, an aryl oxide, a thiol, a halide,an aliphatic amine, or an aromatic amine.
 22. The antiseptic compositionof claim 21, wherein the basic reagent is a guanidium compound.
 23. Theantiseptic composition of claim 22, wherein the guanidium compound ischlorhexidine.
 24. The antiseptic composition of claim 22, wherein theguanidium compound is alexidine.
 25. The antiseptic composition of claim22, wherein the guanidium compound is hexamidine.
 26. The antisepticcomposition of claim 21, wherein the basic reagent is a bipyridine. 27.The antiseptic composition of claim 26, wherein the bipyridine isoctenidine.
 28. The antiseptic composition of claim 21, wherein thebasic reagent is a phenoxide antiseptic.
 29. The antiseptic compositionof claim 28, wherein the phenoxide antiseptic is clofoctol.
 30. Theantiseptic composition of claim 28, wherein the phenoxide antiseptic ischloroxylenol.
 31. The antiseptic composition of claim 28, wherein thephenoxide antiseptic is triclosan.
 32. An antiseptic compound comprisinga basic reagent bound to a dye.
 33. The antiseptic compound of claim 32,wherein the basic reagent and the dye are bound ionically.
 34. Theantiseptic compound of claim 32, wherein the basic reagent and the dyeare bound covalently.
 35. The antiseptic compound of claim 32, whereinthe composition is gendine, genlenol, genlosan, or genfoctol.
 36. Theantiseptic compound of claim 32, further defined by its ability toimpregnate and/or coat a surface.
 37. The antiseptic compound of claim36, wherein the surface is composed of a polymer.
 38. The antisepticcompound of claim 37, wherein the polymer is polyvinyl chloride,polyurethane, polyethylene, silastic elastomers,polytetrafluoroethylene, dacron, collodion, carboethane or nylon. 39.The antiseptic compound of claim 36, wherein said surface is composed ofsilicone.
 40. The antiseptic compound of claim 36, wherein said surfaceis a silk suture.
 41. The antiseptic compound of claim 36, wherein thesurface is an organic surface.
 42. The antiseptic compound of claim 41,wherein the organic surface is skin.
 43. The antiseptic compound ofclaim 41, wherein the organic surface is a mucosal surface.
 44. Theantiseptic compound of claim 41, wherein the organic surface is a wound.45. The antiseptic compound of claim 36, wherein the surface is aninorganic surface.
 46. The antiseptic compound of claim 45, wherein theinorganic surface is a floor.
 47. The antiseptic compound of claim 45,wherein the inorganic surface is a table-top.
 48. The antisepticcompound of claim 45, wherein the inorganic surface is a counter-top.49. The antiseptic compound of claim 45, wherein the inorganic surfaceis the surface of a hospital equipment.
 50. The antiseptic compound ofclaim 45, wherein the inorganic surface is a wheelchair surface.
 51. Amedical device coated with a basic reagent and a dye.
 52. The medicaldevice of claim 50, wherein a basic reagent and a dye are bonded. 53.The medical device of claim 52 wherein the basic reagent and the dye arebound ionically.
 54. The medical device of claim 52, wherein the basicreagent and the dye are bound covalently.
 55. The medical device ofclaim 52, further selected from the group comprising an endotrachealtube, a vascular catheter, an urinary catheter, a nephrostomy tube, abiliary stent, a peritoneal catheter, an epidural catheter, a centralnervous system catheter, an orthopedic device, a prosthetic valve, and amedical implant.
 56. The medical device of claim 55, wherein saidvascular catheter is a central venous catheter, an arterial line, anpulmonary artery catheter, and a peripheral venous catheter.
 57. Themedical device of claim 55, wherein said central nervous system catheteris a intraventricular shunt.
 58. A method for coating a medical devicewith an antiseptic composition comprising: a) immersing said medicaldevice in a solvent comprising a basic reagent and a dye. b) drying thedevice; and c) washing off excessive composition.
 59. The method ofclaim 58, wherein the solvent comprises methylene chloride, methanol, ora combination thereof.
 60. A method for preventing nosocomial infectionsin a subject comprising coating a medical device that the subject isrequired to use with a composition comprising a basic reagent and to adye.
 61. The method of claim 60, wherein said subject is human.
 62. Themethod of claim 60, wherein said nosocomial infection is pneumonia,bacteremia, fungimia, candidemia, a urinary tract infection, acatheter-exit site infection, and a surgical wound infection.
 63. Themethod of claim 60, wherein said nosocomial infection is caused by abacterium.
 64. The method of claim 63, wherein said bacterium is aresistant bacterium.
 65. The method of claim 64, wherein said resistantbacterium is selected from a group comprising methicillin-resistantstaphylococci, vancomycin-resistant enterococci, and resistantPseudomonas aeruginosa.
 66. The method of claim 60, wherein saidnosocomial infection is caused by a fungus.
 67. The method of claim 66,wherein said fungus is a resistant fungus.
 68. The method of claim 67,wherein said resistant fungus belongs to Candida species.
 69. A methodfor disinfecting and/or sterilizing a surface comprising applying acomposition comprising a basic reagent and a dye of claim 1 to thesurface.
 70. The method of claim 69, wherein the surface is an organicsurface.
 71. The method of claim 70, wherein the organic surface isselected from a group comprising, skin, a mucosal surface, and a woundsurface.
 72. The method of claim 69, wherein the surface is an inorganicsurface.
 73. The method of claim 72, wherein the inorganic surface isselected from a group comprising a floor, a table-top, a counter-top,hospital equipment, a wheel chair, gauze, cotton.
 74. A method fordisinfecting and/or sterilizing a fluid comprising adding a compositioncomprising a basic reagent and a dye of claim 1 into the fluid.
 75. Themethod of claim 74, wherein said fluid is water.
 76. The method of claim74 wherein said fluid is a metal working fluid.
 77. The method of claim74, wherein said fluid is petroleum.
 78. A method for preserving asubstance comprising applying a composition comprising a basic reagentand a dye on the substance.
 79. The method of claim 78, wherein thesubstance is selected from the group comprising wood, paint, plastic andpaper.